LAUSR

Abstract BACKGROUND Sitobion avenae (F.) and Rhopalosiphum padi (L.) are harmful pests of wheat [Triticum aestivum (L.)]. No genetic resistance against the aphids has been identified in commercial wheat varieties and resistance phenotyping can be time‐consuming and laborious. Here, we tested a high‐throughput phenotyping method to screen 29 commercial winter wheat varieties for alate antixenosis and antibiosis. We validated this method using comprehensive behavioural analyses, including alate attraction to volatile organic compounds (VOCs) and a feeding bioassay using an electrical penetration graph (EPG), subsequently highlighting possible sources of resistance. RESULTS We observed differences in alate behaviour upon assessing alate settlement on wheat seedlings and attraction towards VOCs, revealing the importance of visual and early post‐alighting cues for alate host selection. Aphid settlement was four times higher on the most preferred variety than on the least preferred variety. Using an EPG bioassay, we identified phloem feeding and stylet derailment parameters linked to resistance. We found antibiosis assessment on detached leaves to be an inadequate screen because it produced results inconsistent with intact leaves assessment. Alate and nymph mortality were identified as key traits signifying antibiosis, showing significant positive relationships with alate reproduction and nymph mean relative growth rate. CONCLUSIONS Overall, antixenosis and antibiosis varietal responses were consistent for both aphid species. Alate settlement on wheat seedlings was a more efficient antixenosis screen than an olfactometer assay using VOCs. In addition to assessing alate and nymph survival for antibiosis, this allows for more rapid phenotyping of large numbers of genotypes to identify novel aphid resistance genes for varietal improvement. © 2024 The Author(s). Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.